Project Summary For HLS17-11, our overall strategy for Hesperos is to utilize microphysiological systems in combination with functional readouts to establish platforms capable of sophisticated analysis of chemicals and drug candidates for toxicity and efficacy during pre-clinical testing, with initial emphasis on predictive toxicity. This is a service based company and is developing low-cost in vitro systems utilizing a novel ?pumpless? microphysiological platform described in US Patent 8,748,180B2. The commercialization potential of our system has been validated by the recent award of a Phase II SBIR to apply advanced manufacturing techniques to increase output and lower cost of production. The pumpless integrated system, using a rocking motion to pump the serum-free cellular medium, reduces the complexity and cost of the fluidic circuit design and simplifies set-up and operation of the device. Hickman has developed microelectrode arrays and cantilever systems that are integrated on chip that allows for noninvasive electronic and mechanical readouts. We have detailed an in vitro cardiac system where the two main components of function, electrical conduction and muscle force, have been reproduced in vitro (Stancescu et al. 2015). The independent measurement of these two key variables allows a detailed description of a compounds effect on overall cardiac function and is currently being used under contract by multiple companies. Because we can measure these functional outputs independently, we can also use these readouts to give ideas on mechanism of action of a compound. Two different sources of cardiomyocytes were used in these studies, iPSCs that are composed of primary cardiomyocytes and cardiomyocytes from embryonic stem cells that contained the three major cell types from the heart. The cardiac cells were shown to reach some aspects of functional maturation as primarily evidenced by patch clamp electrophysiological measurements indicating resting membrane potentials of -85 mV or better. We will adapt this chip-based system to create a microfluidic-based cardiac ischemia model. This cardiac organ-on-a-chip platform will be validated by screening compounds that can restore function after ischemia in this Phase I SBIR. In follow on grants we will look at mechanisms of tissue restoration, screen for cardioprotective compounds as well, and extend the acute experiments to include chronic measurements. A microphysiological system will be developed with functional readouts for cardiac electrical and mechanical function, fitted with environmental sensors, and integrated with an environmental chamber for inducing ischemia. We will first optimize and validate environmental conditions and protocols for inducing and measuring cardiac ischemic damage, followed by validation with ischemia drugs with published in vivo results. The uniqueness will be the combination of Hickman?s functional modules with Shuler?s innovative low-cost ?pumpless? system. Our team contains all of the skill sets required to construct, evaluate and commercialize the integrated system and associated components to achieve the goals.